摘要
目的 探讨细胞周期素D1（ cyclinD1）在上皮性卵巢癌3AO细胞中的表达,分析其表达与3AO细胞生长、增殖的关系.方法 将人正常卵巢上皮细胞及卵巢癌3AO细胞进行体外培养,用逆转录酶多聚酶链式反应（RT-PCR）及流式细胞术（FCM）方法检测受不同浓度顺铂作用前后,细胞中cyclinD1基因及蛋白的表达情况,并观察细胞凋亡及细胞周期的变化.结果 人卵巢上皮细胞中未检测到cyclinD1基因及蛋白的表达,而3AO细胞中存在cyclinD1基因的异常扩增及过表达；受顺铂作用后,3AO细胞琼脂糖凝胶电泳显示cyclinD1基因的mRNA显带由明变暗,逐渐变模糊,并且显带变暗程度与顺铂细胞作用浓度呈正比.流式细胞术直标单抗分析,3AO细胞中cyclinD1蛋白呈阳性过表达；受顺铂作用前后,cyclinD1蛋白表达水平不同,平均值分别是105.9、15.42、8.59;细胞凋亡率不同,分别是0.63％,9.08％,27.41％；细胞周期发生了改变,细胞增殖指数分别是38.83％,44.54％,37.31％,差异有统计学意义（P＜0.01）.结论 卵巢癌3AO细胞中存在cyclinD1基因的异常扩增及过表达,cyclinD1基因及蛋白的阳性表达下降程度与顺铂细胞作用浓度呈正比；cyclinD1的表达与卵巢癌细胞的生长及增殖活性密切相关.
Objective To explore.the expression of cyclinD1 in ovarian carcinoma cell 3AO and analyze its relationship with the proliferation of ovarian cancer cell.Methods Human ovarian epithelium and ovarian cancer cells 3AO were cultured in vito.CyclinD1 genes and proteins were detected by reverse transcription-polymerase chain reaction （RT-PCR） and flow cytometry before and after the activation of cell 3AO by cis-platinum. Also the cell activity and cell cycle were observed. Results Abnormal gene amplification and over-expressions of cyclinD1 were found in ovarian cancer cell while the expression of cyclinD1 was negative in normal ovarian epithelium.Under cis-platinum,different expressions of cyclinD1 genes were found in 3AO by RT-PCR.The higher the concentrations of cisplatin,the lower expressions of cyclinD1 genes.By flow cytometry,it was also found that there were lower expressions of cyclinD1 protein in 3AO under cisplatin than without it. With the rising concentrations of cisplatin,the low expressions of cyclinD1 protein in 3AO were detected.The mean numbers were 105.9,15.42 and 8.59,the cell apoptotic rates 0.63%,9.08% and 27.41% and the proliferation index （PI） numbers 38.83%,44.54%,37.31%.The differences were statistically significant （P 〈 0.01 ）.Conclusion The up-regulation of cyclinD1 is detected in ovarian cancer cell.A positive correlation is found between the lower expressions of cyclinD1 and the concentration of cicplatin.There is a close relationship between the expressions of cyclinD1 and ovarian cancer cell activity.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2012年第5期351-353,共3页
National Medical Journal of China
基金
山东省自然科学基金（Y200LC03）
